Publications by authors named "Jih-Gaw Lin"

42 Publications

Survival strategies of ammonia-oxidizing archaea (AOA) in a full-scale WWTP treating mixed landfill leachate containing copper ions and operating at low-intensity of aeration.

Water Res 2021 Mar 30;191:116798. Epub 2020 Dec 30.

Environmental Engineering, Guangdong Technion - Israel Institute of Technology, 241 Daxue Road, Shantou, Guangdong 515063, China; Southern Laboratory of Ocean Science and Engineering, Zhuhai, Guangdong, China. Electronic address:

Recent studies indicate that ammonia-oxidizing archaea (AOA) may play an important role in nitrogen removal by wastewater treatment plants (WWTPs). However, our knowledge of the mechanisms employed by AOA for growth and survival in full-scale WWTPs is still limited. Here, metagenomic and metatranscriptomic analyses combined with a laboratory cultivation experiment revealed that three active AOAs (WS9, WS192, and WS208) belonging to family Nitrososphaeraceae were active in the deep oxidation ditch (DOD) of a full-scale WWTP treating landfill leachate, which is configured with three continuous aerobic-anoxic (OA) modules with low-intensity aeration (≤ 1.5 mg/L). AOA coexisted with AOB and complete ammonia oxidizers (Comammox), while the ammonia-oxidizing microbial (AOM) community was unexpectedly dominated by the novel AOA strain WS9. The low aeration, long retention time, and relatively high inputs of ammonium and copper might be responsible for the survival of AOA over AOB and Comammox, while the dominance of WS9, specifically may be enhanced by substrate preference and uniquely encoded retention strategies. The urease-negative WS9 is specifically adapted for ammonia acquisition as evidenced by the high expression of an ammonium transporter, whereas two metabolically versatile urease-positive AOA strains (WS192 and WS208) can likely supplement ammonia needs with urea. This study provides important information for the survival and application of the eutrophic Nitrososphaeraceae AOA and advances our understanding of archaea-dominated ammonia oxidation in a full-scale wastewater treatment system.
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http://dx.doi.org/10.1016/j.watres.2020.116798DOI Listing
March 2021

Specific and effective detection of anammox bacteria using PCR primers targeting the 16S rRNA gene and functional genes.

Sci Total Environ 2020 Sep 15;734:139387. Epub 2020 May 15.

School of Food and Biotechnology, Guangdong Industry Polytechnic, Guangzhou, Guangdong 510300, People's Republic of China; Environmental Engineering, Guangdong Technion Israel Institute of Technology, 241 Daxue Road, Shantou, Guangdong 515063, People's Republic of China. Electronic address:

Anaerobic ammonium-oxidizing (anammox) bacteria play an important role in the nitrogen cycle by coupling ammonium and nitrite to produce dinitrogen gas (N). Polymerase chain reaction (PCR) is a fast, simple, and sensitive method that is widely used to assess the diversity, abundance, and activity of the slow-growing bacteria. In this review, we summarize and evaluate the wide variety of PCR primers targeting the 16S rRNA gene and functional genes (hzo, nir, and hzs) of anammox bacteria for their effectiveness and efficiencies in detecting this group of bacteria in different sample types. Furthermore, the efficiencies of different universal high-throughput sequencing 16S rRNA gene primers in anammox bacteria investigations were also evaluated to provide a reference for primer selection. Based on our in silico evaluation results, none of the 16S rRNA gene primers could recover all of the known anammox bacteria, but multiple hzo and hzs gene primers could accomplish this task. However, uncertain copies (1-3 copies) of hzo genes were identified in the genomes, and the hydrazine oxidation reaction catalyzed by hydrazine oxidoreductases (HZOs) can also be catalyzed by other hydroxylamine oxidoreductases (HAOs) in anammox bacteria, which can potentially result in large deviations in hzo-based qPCR and RT-qPCR analyses and results. Therefore, the use of optimal primers targeting unique hzs genes are recommended, although the efficiencies of these newly designed primers need further verification in practical applications. This article provides comprehensive information for the effective and specific detection of anammox bacteria using specific primers targeting the 16S rRNA gene and functional genes and serves as a basis for future high-quality primer design.
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http://dx.doi.org/10.1016/j.scitotenv.2020.139387DOI Listing
September 2020

Nanoarchitectured structure and population dynamics of anaerobic ammonium oxidizing (anammox) bacteria in a wastewater treatment plant.

J Hazard Mater 2020 09 19;396:122714. Epub 2020 Apr 19.

Institute of Environmental Engineering, National Chiao Tung University, 1001 University Road, Hsinchu City 30010, Taiwan.

Studies on microbial community and population dynamics are essential for the successful development, monitoring and operation of biological wastewater treatment systems. Especially for novel or sustainable systems such as the anaerobic ammonium oxidizing (anammox) process that are not yet well explored. Here we collected granular microbial sludge samples and investigated a community of anammox bacteria over a period of four years, divided into eight stages in a full scale simultaneous partial nitrification, anammox and denitrification (SNAD) process for treating landfill leachate. Specific qPCR primers were designed to target and quantify the two most abundant anammox species, Candidatus Kuenenia stuttgartiensis (KS) and Candidatus Brocadia anammoxidans (BA). The two species were monitored and could explain the dynamic shift of the anammox community corresponding to the operating conditions. Using the newly designed KS-specific primer (KSqF3/KSqR3) and BA-specific primer (BAqF/BAqR), we estimated the amounts of KS and BA to be in the range of 6.2 × 10 to 5.9 × 10 and 1.1 × 10 to 4.1 × 10 copies μg DNA, respectively. KS was found to be the dominant species in all anammox granules studied and played an important role in the formation of granules. The KS/BA ratio was positively correlated to the size of granules in the reactor and ammonia nitrogen removal efficiency of the treatment plant.
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http://dx.doi.org/10.1016/j.jhazmat.2020.122714DOI Listing
September 2020

Activity and Metabolic Versatility of Complete Ammonia Oxidizers in Full-Scale Wastewater Treatment Systems.

mBio 2020 03 17;11(2). Epub 2020 Mar 17.

Laboratory of Environmental Microbiology and Toxicology, School of Biological Sciences, The University of Hong Kong, Hong Kong SAR, Hong Kong, People's Republic of China

The recent discovery of complete ammonia oxidizers (comammox) contradicts the paradigm that chemolithoautotrophic nitrification is always catalyzed by two different microorganisms. However, our knowledge of the survival strategies of comammox in complex ecosystems, such as full-scale wastewater treatment plants (WWTPs), remains limited. Analyses of genomes and transcriptomes of four comammox organisms from two full-scale WWTPs revealed that comammox were active and showed a surprisingly high metabolic versatility. A gene cluster for the utilization of urea and a gene encoding cyanase suggest that comammox may use diverse organic nitrogen compounds in addition to free ammonia as the substrates. The comammox organisms also encoded the genomic potential for multiple alternative energy metabolisms, including respiration with hydrogen, formate, and sulfite as electron donors. Pathways for the biosynthesis and degradation of polyphosphate, glycogen, and polyhydroxyalkanoates as intracellular storage compounds likely help comammox survive unfavorable conditions and facilitate switches between lifestyles in fluctuating environments. One of the comammox strains acquired from the anaerobic tank encoded and transcribed genes involved in homoacetate fermentation or in the utilization of exogenous acetate, both pathways being unexpected in a nitrifying bacterium. Surprisingly, this strain also encoded a respiratory nitrate reductase which has not yet been found in any other genome and might confer a selective advantage to this strain over other strains in anoxic conditions. The discovery of comammox in the genus changes our perception of nitrification. However, genomes of comammox organisms have not been acquired from full-scale WWTPs, and very little is known about their survival strategies and potential metabolisms in complex wastewater treatment systems. Here, four comammox metagenome-assembled genomes and metatranscriptomic data sets were retrieved from two full-scale WWTPs. Their impressive and-among nitrifiers-unsurpassed ecophysiological versatility could make comammox an interesting target for optimizing nitrification in current and future bioreactor configurations.
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http://dx.doi.org/10.1128/mBio.03175-19DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7078480PMC
March 2020

Complex microbial nitrogen-cycling networks in three distinct anammox-inoculated wastewater treatment systems.

Water Res 2020 Jan 30;168:115142. Epub 2019 Sep 30.

Laboratory of Environmental Microbiology and Toxicology, School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong Kong, China. Electronic address:

Microbial nitrogen removal mediated by anaerobic ammonium oxidation (anammox) are cost-effective, yet it is time-consuming to accumulate the slow-growing anammox bacteria in conventional wastewater treatment plants (WWTPs). Inoculation of anammox enriched pellets is an effective way to establish anammox and achieve shortcut nitrogen removal in full-scale WWTPs. However, little is known about the complex microbial nitrogen-cycling networks in these anammox-inoculated WWTPs. Here, we applied metagenomic and metatranscriptomic tools to study the microbial nitrogen removal in three conventional WWTPs, which have been inoculated exogenous anammox pellets, representing partial-nitrification anammox (PNA) and nitrification-denitrification nitrogen removal processes. In the PNA system of Bali (BL), ammonia was partially oxidized by ammonia-oxidizing bacteria (AOB) Nitrosomonas and the oxidized nitrite and the remaining ammonium were directly converted to N by anammox bacteria Ca. Brocadia and Ca. Kuenenia. In the nitrification-denitrification system of Wenshan (WS), ammonia-oxidizing archaea (AOA) Thaumarchaeota unexpectedly dominated the nitrifying community in the presence of AOB Nitrosomonas. Meanwhile, the biomass yield of Ca. Brocadia was likely inhibited by the high biodegradable organic compound input and limited by substrate competitions from AOA, AOB, complete ammonia oxidizers (comammox) Nitrospira, nitrite-oxidizing bacteria (NOB) Nitrospira, and heterotrophic denitrifiers. Unexpectedly, comammox Nitrospira was the predominant nitrifier in the presence of AOB Nitrosomonas in the organic carbon-rich nitrification-denitrification system of Linkou (LK). These results clearly showed that distinct active groups were working in concert for an effective nitrogen removal in different WWTPs. This study confirmed the feasibility of anammox application in ammonium-rich systems by direct inoculation of the exogenous anammox pellets and improved our understanding of microbial nitrogen cycling in anammox-driven conventional WWTPs from both physiochemical and omics perspectives.
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http://dx.doi.org/10.1016/j.watres.2019.115142DOI Listing
January 2020

A xylan-degrading thermophilic and obligate anaerobe Xylanivirga thermophila gen. nov., sp. nov., isolated from an anammox dominant wastewater treatment plant, and proposal of Xylanivirgaceae fam. nov.

Anaerobe 2020 Feb 18;61:102075. Epub 2019 Jul 18.

State Key Laboratory of Biocontrol and Guangdong Provincial Key Laboratory of Plant Resources, School of Life Sciences, Sun Yat-Sen University, Guangzhou, 510275, PR China; Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences, Ürűmqi, 830011, PR China. Electronic address:

In our search for novel anaerobes with potential carbohydrate polymers degrading activity, we have isolated a xylan-degrading bacterial strain SYSU GA17129 from an anammox bacteria dominant wastewater treatment plant. Phylogenetic analysis of the 16S rRNA gene sequence indicated the strain SYSU GA17129 belong to the order Clostridiales and shared highest sequence identity with Caldicoprobacter faecalis DSM 20678 (89.9%). The strain was thermophilic, obligately anaerobic, non-motile and rod shaped. Optimum growth of the strain was observed at 45 °C, pH 8.0 and in the presence of 0.5% NaCl (w/v). The chemotaxonomic features of the strain SYSU GA17129 comprised of C FAME, iso-C FAME and C FAME as the major fatty acids (>10%), diphosphatidylglycerol, phosphatidylinositol mannoside, an unidentified phospholipid, three unidentified polar lipids and two unidentified glycolipids as its polar lipids, and meso-diaminopimelic acid (meso-DAP) as the diamino acid in peptidoglycan. The G + C content of the genomic DNA was 35.9%. The strain could be distinguished from other defined families within the order Clostridiales by the differences in phenotypic and physiological characteristics, distinct phylogenetic lineage in 16S rRNA gene- and genome-based phylogenies and low genomic relatedness index. Based on these distinguishing properties, strain SYSU GA17129 is proposed to represent a new species of a new genus Xylanivirga thermophila gen. nov., sp. nov., within a new family Xylanivirgaceae fam. nov. The type species of the new taxon is SYSU GA17129 (=KCTC 15754 = CGMCC 1.5282). This strain is characterized within the order Clostridiales, class Clostridia of the phylum Firmicutes.
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http://dx.doi.org/10.1016/j.anaerobe.2019.102075DOI Listing
February 2020

The short- and long-term inhibitory effects of Fe (II) on anaerobic ammonium oxidizing (anammox) process.

Water Sci Technol 2019 May;79(10):1860-1867

Faculty of Engineering and Green Technology, University Tunku Abdul Rahman, Kampar, Malaysia E-mail:

The application of the anammox process has great potential in treating nitrogen-rich wastewater. The presence of Fe (II) is expected to affect the growth and activity of anammox bacteria. Short-term (acute) and long-term effects (chronic) of Fe (II) on anammox activity were investigated. In the short-term study, results demonstrated that the optimum concentration of Fe (II) that could be added to anammox is 0.08 mM, at which specific anammox activity (SAA) improved by 60% compared to the control assay, 0.00 mM. The inhibition concentration, IC, of Fe (II) was found to be 0.192 mM. Kinetics of anammox specific growth rate were estimated based on results of the batch test and evaluated with Han-Levenspiel's substrate inhibition kinetics model. The optimum concentration and IC of Fe (II) predicted by the Han-Levenspiel model was similar to the batch test, with values of 0.07 mM and 0.20 mM, respectively. The long-term effect of Fe (II) on the performance of a sequencing batch reactor (SBR) was evaluated. Results showed that an appropriate Fe (II) addition enhanced anammox activity, achieving 85% NH-N and 96% NO-N removal efficiency when 0.08 mM of Fe (II) was added. Quantitative polymerase chain reaction (qPCR) was adopted to detect and identify the anammox bacteria.
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http://dx.doi.org/10.2166/wst.2019.188DOI Listing
May 2019

Metagenomic and metatranscriptomic analyses reveal activity and hosts of antibiotic resistance genes in activated sludge.

Environ Int 2019 08 23;129:208-220. Epub 2019 May 23.

Institute for Advanced Study, Shenzhen University, Shenzhen, Guangdong, PR China. Electronic address:

Wastewater treatment plants (WWTPs) are a source and reservoir for subsequent spread of various antibiotic resistance genes (ARGs). However, little is known about the activity and hosts of ARGs in WWTPs. Here, we utilized both metagenomic and metatranscriptomic approaches to comprehensively reveal the diversity, abundance, expression and hosts of ARGs in activated sludge (AS) from three conventional WWTPs in Taiwan. Based on deep sequencing data and a custom-made ARG database, a total of 360 ARGs associated with 24 classes of antibiotics were identified from the three AS metagenomes, with an abundance range of 7.06 × 10-1.20 × 10 copies of ARG/copy of 16S rRNA gene. Differential coverage binning analysis revealed that >22 bacterial phyla were the putative hosts of the identified ARGs. Surprisingly, genus Mycobacterium and family Burkholderiaceae were observed as multi-drug resistant harboring 14 and 50 ARGs. Metatranscriptome analysis showed 65.8% of the identified ARGs were being expressed, highlighting that ARGs were not only present, but also transcriptionally active in AS. Remarkably, 110 identified ARGs were annotated as plasmid-associated and displayed a close to two-fold increased likelihood of being transcriptionally expressed compared to those ARGs found exclusively within bacterial chromosomes. Further analysis showed the transcript abundance of aminoglycoside, sulfonamide, and tetracycline resistance genes was mainly contributed by plasmid-borne ARGs. Our approach allowed us to specifically link ARGs to their transcripts and genetic context, providing a comprehensive insight into the prevalence, expression and hosts of ARGs in AS. Overall, results of this study enhance our understanding of the distribution and dissemination of ARGs in WWTPs, which benefits environmental risk assessment and management of ARB and ARGs.
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http://dx.doi.org/10.1016/j.envint.2019.05.036DOI Listing
August 2019

Assessment of molecular detection of anaerobic ammonium-oxidizing (anammox) bacteria in different environmental samples using PCR primers based on 16S rRNA and functional genes.

Appl Microbiol Biotechnol 2017 Oct 20;101(20):7689-7702. Epub 2017 Sep 20.

Laboratory of Environmental Microbiology and Toxicology, School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong Kong, People's Republic of China.

Eleven published PCR primer sets for detecting genes encoding 16S ribosomal RNA (rRNA), hydrazine oxidoreductase (HZO), cytochrome cd -containing nitrite reductase (NirS), and hydrazine synthase subunit A (HzsA) of anaerobic ammonium-oxidizing (anammox) bacteria were assessed for the diversity and abundance of anammox bacteria in samples of three environments: wastewater treatment plant (WWTP), wetland of Mai Po Nature Reserve (MP), and the South China Sea (SCS). Consistent phylogenetic results of three biomarkers (16S rRNA, hzo, and hzsA) of anammox bacteria were obtained from all samples. WWTP had the lowest diversity with Candidatus Kuenenia dominating while the SCS was dominated by Candidatus Scalindua. MP showed the highest diversity of anammox bacteria including C. Scalindua, C. Kuenenia, and Candidatus Brocadia. Comparing different primer sets, no significant differences in specificity for 16S rRNA gene could be distinguished. Primer set CL1 showed relatively high efficiency in detecting the anammox bacterium hzo gene from all samples, while CL2 showed greater selectivity for WWTP samples. The recently reported primer sets of the hzsA gene resulted in high efficiencies in detecting anammox bacteria while nirS primer sets were more selective for specific samples. Results collectively indicate that the distribution of anammox bacteria is niche-specific within different ecosystems and primer specificity may cause biases on the diversity detected.
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http://dx.doi.org/10.1007/s00253-017-8502-3DOI Listing
October 2017

Current advances in molecular methods for detection of nitrite-dependent anaerobic methane oxidizing bacteria in natural environments.

Appl Microbiol Biotechnol 2016 Dec 20;100(23):9845-9860. Epub 2016 Oct 20.

Laboratory of Environmental Microbiology and Toxicology, School of Biological Sciences, Faculty of Science, The University of Hong Kong, Pokfulam Road, Hong Kong, SAR, People's Republic of China.

Nitrite-dependent anaerobic methane oxidation (n-damo) process uniquely links microbial nitrogen and carbon cycles. Research on n-damo bacteria progresses quickly with experimental evidences through enrichment cultures. Polymerase chain reaction (PCR)-based methods for detecting them in various natural ecosystems and engineered systems play a very important role in the discovery of their distribution, abundance, and biodiversity in the ecosystems. Important characteristics of n-damo enrichments were obtained and their key significance in microbial nitrogen and carbon cycles was investigated. The molecular methods currently used in detecting n-damo bacteria were comprehensively reviewed and discussed for their strengths and limitations in applications with a wide range of samples. The pmoA gene-based PCR primers for n-damo bacterial detection were evaluated and, in particular, several incorrectly stated PCR primer nucleotide sequences in the published papers were also pointed out to allow correct applications of the PCR primers in current and future investigations. Furthermore, this review also offers the future perspectives of n-damo bacteria based on current information and methods available for a better acquisition of new knowledge about this group of bacteria.
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http://dx.doi.org/10.1007/s00253-016-7853-5DOI Listing
December 2016

Biodegradation of tetramethylammonium hydroxide (TMAH) in completely autotrophic nitrogen removal over nitrite (CANON) process.

Bioresour Technol 2016 Jun 12;210:88-93. Epub 2016 Feb 12.

Institute of Environmental Engineering, National Chiao Tung University, 1001 University Road, Hsinchu 30010, Taiwan. Electronic address:

This study conducted a completely autotrophic nitrogen removal over nitrite (CANON) process in a continuous anoxic upflow bioreactor to treat synthetic wastewater with TMAH (tetramethylammonium hydroxide) ranging from 200 to 1000mg/L. The intermediates were analyzed for understanding the metabolic pathway of TMAH biodegradation in CANON process. In addition, (15)N-labeled TMAH was used as the substrate in a batch anoxic bioreactor to confirm that TMAH was converted to nitrogen gas in CANON process. The results indicated that TMAH was almost completely biodegraded in CANON system at different influent TMAH concentrations of 200, 500, and 1000mg/L. The average removal efficiencies of total nitrogen were higher than 90% during the experiments. Trimethylamine (TMA) and methylamine (MA) were found to be the main biodegradation intermediates of TMAH in CANON process. The production of nitrogen gas with (15)N-labeled during the batch anaerobic bioreactor indicated that CANON process successfully converted TMAH into nitrogen gas.
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http://dx.doi.org/10.1016/j.biortech.2016.01.127DOI Listing
June 2016

Start-up of simultaneous partial nitrification, anammox and denitrification (SNAD) process in sequencing batch biofilm reactor using novel biomass carriers.

Bioresour Technol 2015 Aug 20;190:480-6. Epub 2015 Feb 20.

Institute of Environmental Engineering, National Chiao Tung University, Hsinchu 30010, Taiwan. Electronic address:

Simultaneous partial nitrification, anammox and denitrification (SNAD) process was started-up in a 2.5L sequencing batch biofilm reactor (SBBR) using novel biomass carriers. The SNAD process took only 51d for start-up at nitrogen loading rate (NLR) and organic loading rate (OLR) of 120 and 60g/m(3)-d, respectively. Long-term stable operation of SNAD process was observed at NLR and OLR of 360 and 180g/m(3)-d with average total nitrogen and COD removal efficiencies of >88% and >90%, respectively. The values of conversion ratio [Formula: see text] remained below 0.11 after the start-up period, which further confirmed the long-term stability of SNAD process. Results of polymerase chain reaction (PCR), qualitative PCR, and scanning electron microscopic (SEM) analysis of sludge samples confirmed the co-existence and enrichment of AOB, anammox bacteria and denitrifying bacteria in the reactor and biofilm formation on to the carriers.
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http://dx.doi.org/10.1016/j.biortech.2015.02.064DOI Listing
August 2015

Effect of zinc on anammox activity and performance of simultaneous partial nitrification, anammox and denitrification (SNAD) process.

Bioresour Technol 2014 Aug 18;165:105-10. Epub 2014 Apr 18.

Institute of Environmental Engineering, National Chiao Tung University, Hsinchu 30010, Taiwan. Electronic address:

In the present study, short-term effects of zinc on anammox activities and long-term effect of zinc on the performance of simultaneous partial nitrification, anammox and denitrification (SNAD) process were evaluated. The anammox activity decreased with increasing zinc concentration and exposure time in short-term tests. The IC50 value of zinc was found to be 6.9mg/L. However, the presence of zinc (<10mg/L) in wastewater stimulated the microbial activities and nitrogen removal performance of SNAD process in sequencing batch biofilm reactor (SBBR). At first, inhibition of SNAD process was observed when influent zinc concentration increased to 20mg/L. The system recovered immediately, suggesting the acclimatization of microbial communities of SNAD process. The results showed that SBBR was well acclimatized under high zinc concentration (50-100mg/L) achieving 98% NH4(+)-N, 96% TN and 87% COD removal efficiencies.
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http://dx.doi.org/10.1016/j.biortech.2014.04.034DOI Listing
August 2014

Removal of pharmaceuticals and organic matter from municipal wastewater using two-stage anaerobic fluidized membrane bioreactor.

Bioresour Technol 2014 Aug 21;165:42-9. Epub 2014 Mar 21.

Institute of Environmental Engineering, National Chiao Tung University, 1001 University Road, Hsinchu City 30010, Taiwan. Electronic address:

The aim of present study was to treat municipal wastewater in two-stage anaerobic fluidized membrane bioreactor (AFMBR) (anaerobic fluidized bed reactor (AFBR) followed by AFMBR) using granular activated carbon (GAC) as carrier medium in both stages. Approximately 95% COD removal efficiency could be obtained when the two-stage AFMBR was operated at total HRT of 5h (2h for AFBR and 3h for AFMBR) and influent COD concentration of 250mg/L. About 67% COD and 99% TSS removal efficiency could be achieved by the system treating the effluent from primary clarifier of municipal wastewater treatment plant, at HRT of 1.28h and OLR of 5.65kg COD/m(3)d. The system could also effectively remove twenty detected pharmaceuticals in raw wastewaters with removal efficiency in the range of 86-100% except for diclofenac (78%). No other membrane fouling control was required except scouring effect of GAC for flux of 16LMH.
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http://dx.doi.org/10.1016/j.biortech.2014.03.054DOI Listing
August 2014

Study of chemical pretreatment and enzymatic saccharification for producing fermentable sugars from rice straw.

Bioprocess Biosyst Eng 2014 Jul 18;37(7):1337-44. Epub 2013 Dec 18.

Department of Environmental Engineering, National Ilan University, Yilan, 260, Taiwan,

This study evaluated a cost-effective approach for the conversion of rice straw into fermentable sugars. The composition of rice straw pretreated with 1 % sulfuric acid or 1 % sodium hydroxide solution was compared to rice straw with no chemical pretreatment. Enzymatic saccharification experiments on non-pretreated rice straw (NPRS), pretreated rice straw (PRS), and pretreated rice straw with acid hydrolysate (PRSAH) were conducted in a series of batch reactors. The results indicated that pretreating the rice straw with dilute acid and base increased the cellulose content from 38 % to over 50 %. During enzymatic saccharification, straight aliphatic cellulose was hydrolyzed before branched hemicellulose, and glucose was the major hydrolysis product. The glucose yield was 0.52 g glucose/g for NPRS and was comparable to the yields of 0.50 g glucose/g for PRS and 0.58 g glucose/g for PRSAH. The hydrolysis of rice straw to produce glucose can be described by a first-order reaction with a rate constant of 0.0550 d(-1) for NPRS, 0.0653 d(-1) for PRSAH, and 0.0654 d(-1) for PRS. Overall, the production of fermentable sugars from ground rice straw will be more cost effective if the straw is not pretreated with chemicals.
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http://dx.doi.org/10.1007/s00449-013-1106-0DOI Listing
July 2014

A comparison of two 16S rRNA gene-based PCR primer sets in unraveling anammox bacteria from different environmental samples.

Appl Microbiol Biotechnol 2013 Dec 1;97(24):10521-9. Epub 2013 Nov 1.

Laboratory of Environmental Microbiology and Toxicology, School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong Kong SAR, Hong Kong, People's Republic of China.

Two 16S rRNA gene-based PCR primer sets (Brod541F/Amx820R and A438f/A684r) for detecting anammox bacteria were compared using sediments from Mai Po wetlands (MP), the South China Sea (SCS), a freshwater reservoir (R2), and sludge granules from a wastewater treatment plant (A2). By comparing their ability in profiling anammox bacteria, the recovered diversity, community structure, and abundance of anammox bacteria among all these diverse samples indicated that A438f/A684r performed better than Brod541F/Amx820R in retrieving anammox bacteria from these different environmental samples. Five Scalindua subclusters (zhenghei-I, SCS-I, SCS-III, arabica, and brodae) dominated in SCS whereas two Scalindua subclusters (zhenghei-II and wagneri) and one cluster of Kuenenia dominated in MP. R2 showed a higher diversity of anammox bacteria with two new retrieved clusters (R2-New-1 and R2-New-2), which deserves further detailed study. The dominance of Brocadia in sample A2 was supported by both of the primer sets used. Results collectively indicate strongly niche-specific community structures of anammox bacteria in different environments, and A438f/A684r is highly recommended for screening anammox bacteria from various environments when dealing with a collection of samples with diverse physiochemical characteristics.
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http://dx.doi.org/10.1007/s00253-013-5305-zDOI Listing
December 2013

Ambient temperature SNAD process treating anaerobic digester liquor of swine wastewater.

Bioresour Technol 2013 Aug 24;141:191-8. Epub 2013 Feb 24.

Institute of Environmental Engineering, National Chiao Tung University, Hsinchu 30010, Taiwan.

In present study, effluent from anaerobic digestion of swine wastewater was treated by the simultaneous partial nitrification, anaerobic ammonium oxidation and denitrification (SNAD) process using a lab scale 5L sequencing batch reactor (SBR) under ambient temperature. The fluctuation of anaerobic digester liquor quality (COD, 387 ± 145 mg/L; TKN, 662 ± 190 mg/L; NH₄(+)-N, 519 ± 134 mg/L) and temperature created difficulties to develop a stable SNAD process in the SBR (days 1-285). Fed batch feeding strategy was adopted to have a stable condition in the reactor and overcome the negative effects of organic nitrogen. The average total nitrogen, NH₄(+)-N and COD removal efficiencies in the SBR under steady state conditions (days 485-523) were 80%, 96% and 76%, respectively. The results showed that presence of organic nitrogen, mode of feeding and reactor temperature affects the SNAD process.
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http://dx.doi.org/10.1016/j.biortech.2013.02.045DOI Listing
August 2013

Partial nitrification and anammox process: a method for high strength optoelectronic industrial wastewater treatment.

Water Res 2013 Jun 4;47(9):2929-37. Epub 2013 Feb 4.

Institute of Environmental Engineering, National Chiao Tung University, 1001 University Road, Hsinchu City 30010, Taiwan, ROC.

Completely autotrophic nitrogen removal over nitrite (CANON) process was employed in an 18 L sequencing batch reactor (SBR) for treatment of optoelectronic industrial wastewater containing high strength ammonium nitrogen (3712 ± 120 mg NH4(+) - N L(-1)). About 89% of total nitrogen and 98% of NH4(+) - N removal efficiencies were observed at the loading rate of 909 g N m(-3) d(-1) and the HRT of 4 d. A profound variation in the performance of CANON process was experienced at high DO exposure (above 1 mg L(-1)) and high nitrite concentration (above 100 mg L(-1)). Inhibition due to high DO exposure was found to be reversible phenomenon whereas the synergistic inhibition of nitrite, free ammonia and free nitrous acid was irreversible. The fluctuation of reactor temperature between 17 and 37 °C did not affect the performance of CANON system. The CANON process was stably controlled at high nitrogen loading rate for more than one month. The co-existence of aerobic and anaerobic ammonium oxidizing bacteria in the reactor was detected by The PCR analysis. About 5 fold increase in amount of anammox bacteria over a period of 258 days was confirmed from the results of qPCR on day 487.
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http://dx.doi.org/10.1016/j.watres.2013.01.028DOI Listing
June 2013

Evaluation of biobutanol production from non-pretreated rice straw hydrolysate under non-sterile environmental conditions.

Bioresour Technol 2013 May 5;135:262-8. Epub 2012 Nov 5.

Department of Environmental Engineering, National Ilan University, Yilan 260, Taiwan.

The study aims to investigate a cost-effective approach to convert non-pretreated rice straw hydrolysate into biobutanol. The influences of the initial cell concentration and incubation temperature on biobutanol production were evaluated under both sterile and non-sterile conditions. Results indicate that 100% glucose utilization could be achieved for initial cell concentrations greater than 2100 mg/L under both sterile and non-sterile conditions. Regression analyses resolve that under the sterile condition, the maximum butanol productivity of 1.45 g/L/d was projected at 1.96 g/L of cells and 32.3 °C, while the maximum butanol yield of 0.22 g/g was predicted at 2.01 g/L of cells and 26.3 °C. These two maximum values could not be projected by the regression analyses for the non-sterile condition. However, this study confirms that a high initial cell concentration of Clostridium saccharoperbutylacetonicum N1-4 can minimize interference from other microbes so that non-sterile biobutanol production is comparable to sterile biobutanol production.
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http://dx.doi.org/10.1016/j.biortech.2012.10.140DOI Listing
May 2013

Effect of supplementary carbon addition in the treatment of low C/N high-technology industrial wastewater by MBR.

Bioresour Technol 2012 Jun 27;113:148-53. Epub 2011 Dec 27.

Department of Civil Engineering, National Institute of Technology Calicut, Kozhikode, Kerala 673601, India.

The effect of supplementary carbon addition for the treatment of high-technology industrial wastewater in a membrane bioreactor (MBR) was investigated. The MBR was operated for 302 days under different C/N (BOD(L)/NH(4)(+)-N) ratios, i.e. 0.9-1 to 20 days, 1.6-21 to 42 days, 2.9-43 to 82 days, 3.6-83 to 141 days, 4.8-165 to 233 days and 9.3-240 to 302 days. Irrespective of the C/N ratios investigated, SS and BOD(5) removal efficiencies were above 95% and above 80% COD removal efficiency was observed. In addition, complete nitrification was observed throughout the investigation. However, denitrification and total nitrogen removal efficiencies reached their maximum values at the highest C/N ratio (9.3) investigated. Real-time PCR analysis revealed 10 times higher ammonia oxidizing bacteria to total bacteria ratio under the highest C/N ratio condition (9.3) compared to the low C/N ratio condition (1.6).
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http://dx.doi.org/10.1016/j.biortech.2011.12.102DOI Listing
June 2012

Nitrogen removal from opto-electronic wastewater using the simultaneous partial nitrification, anaerobic ammonium oxidation and denitrification (SNAD) process in sequencing batch reactor.

Bioresour Technol 2012 Jun 14;113:225-31. Epub 2011 Dec 14.

Institute of Environmental Engineering, National Chiao Tung University, 1001 University Road, Hsinchu City 30010, Taiwan, ROC.

Simultaneous partial nitrification, anaerobic ammonium oxidation and denitrification (SNAD) system was applied to treat the opto-electronic industrial wastewater in a 2.5L sequencing batch reactor. The characteristics of wastewater were low C/N ratio (≈ 0.2) with 100mg-COD/L and 567 mg NH(4)(+)-N/L. The experiment was carried out over 8 months in six different stages, where nitrogen loading rate was gradually increased from 16 g-N/m(3)d in stage I to 230 g-N/m(3)d in stage VI. The COD and nitrogen removal rates by the SNAD system reached to 28 g COD/m(3)d and 197 g NH(4)(+)-N/m(3)d, respectively in stage VI. These results showed that the SNAD system is suitable to treat wastewater containing high nitrogen pollutants with low COD level. Presence of ammonium oxidizing bacteria and anammox bacteria were confirmed by polymerase chain reaction (PCR). PCR results also indicated that Candidatus Kuenenia stuttgartiensis may be one of the dominant species in the reactor.
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http://dx.doi.org/10.1016/j.biortech.2011.12.004DOI Listing
June 2012

Sono-alkalization pretreatment of sewage sludge containing phthalate acid esters.

J Environ Sci Health A Tox Hazard Subst Environ Eng 2011 ;46(9):980-8

Department of Environmental Engineering and Health, Yuanpei University, Hsinchu, Taiwan.

This work experimentally elucidates the efficiencies of sono-alkalization treatment on municipal sewage sludge. The total solids (TS) concentration of the sewage sludge was pre-adjusted at 29.7 g/L. Two parameters such as sodium hydroxide (NaOH) dosage and sonication time were considered by the central composite design (CCD) program to investigate the effect on the degradation of phthalate acid esters (PAEs) and solubilization of soluble chemical oxygen demand (SCOD). The mean concentrations of dibutyl phthalate (DBP), di-(2-ethylhexyl) phthalate (DEHP) and butyl benzyl phthalate (BBP) in the sewage sludge were 718, 41 and 8 mg/kg dry weight sludge, respectively. Sono-alkalization process was effective on the degradation of DBP but worthless for DEHP and BBP. Overall degradation of DBP in sewage sludge was estimated to be 100% at the NaOH dosage of 68 m Mand sonication time of 2.2 min. Sono-alkalization was responsible for 6,000 mg/L increase of SCOD based on the decrease of volatile solids in sewage sludge.
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http://dx.doi.org/10.1080/10934529.2011.586261DOI Listing
October 2011

Carbofuran degradation by the application of MW-assisted H₂O₂ process.

J Environ Sci Health B 2011 ;46(4):350-9

Institute of Environmental Engineering, National Chiao Tung University, Hsinchu City, Taiwan.

Carbofuran removal performance of a microwave (MW)-assisted H₂O₂ system under different MW-power levels (300-900 W) was investigated. Batch experiments were conducted at 100 mg/L carbofuran concentration using a modified-MW reactor with 2450 MHz of fixed frequency. As a precursor, control experiments were carried out with H₂O₂ alone, MW alone and conventional heating (CH). A maximum carbofuran removal of 14 % was observed in both H₂O₂ alone and CH systems. On the other hand, only 2 % removal was observed in the MW alone system irrespective of the operation-mode, i.e. continuous or pulsed. The combination of MW and H₂O₂ produced 100 % carbofuran removal in all the MW-assisted experiments. The MW-assisted system operated under continuous-mode and at 750 W has showed rapid carbofuran degradation, i.e. 30 sec, with the highest first-order removal rate constant of 25.82/min. However, 97 % carbon oxygen demand (COD) removal was observed in the same system only after 30 min. On the other hand, 100 % carbofuran removal and 49 % COD removal were observed in the pulsed-mode MW-assisted H₂O₂ system after 10 and 30 min, respectively. Carbofuran mineralization in the system was evidenced by the formation of ammonium and nitrate, and carbofuran intermediates.
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http://dx.doi.org/10.1080/03601234.2011.559894DOI Listing
August 2011

Fast start-up, performance and microbial community in a pilot-scale anammox reactor seeded with exotic mature granules.

Bioresour Technol 2011 Feb 9;102(3):2448-54. Epub 2010 Nov 9.

School of Chemistry and Chemical Engineering, Shandong University, Jinan, China.

The possibility to introduce the exotic anammox sludge to seed the pilot-scale anammox granular reactor and its fast start-up for treating high nitrogen concentration wastewater were evaluated in this study. The reactor was started up successfully in two weeks; in addition, high nitrogen removal was achieved for a long period. Stoichiometry molar ratios of nitrite conversion and nitrate production to ammonium conversion were calculated to be 1.26±0.02:1 and 0.26±0.01:1, respectively. The Stover-Kincannon model which was first applied in granular anammox process indicated that the granular anammox reactor possessed high nitrogen removal potential of 27.8 kg/m(3)/d. The anammox granules in the reactor were characterized via microscope observation and fluorescence in situ hybridization technique. Moreover, the microbial community of the granules was quantified to be composed of 91.4-92.4% anammox bacteria by real-time polymerase chain reaction. This pilot study can elucidate further information for industrial granular anammox application.
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http://dx.doi.org/10.1016/j.biortech.2010.11.006DOI Listing
February 2011

Development of simultaneous partial nitrification, anammox and denitrification (SNAD) process in a sequential batch reactor.

Bioresour Technol 2011 May 12;102(9):5514-9. Epub 2010 Nov 12.

Institute of Environmental Engineering, National Chiao Tung University, 1001 University Road, Hsinchu City 30010, Taiwan, ROC.

Simultaneous partial nitrification, anammox and denitrification (SNAD) process was developed in a sequential batch reactor (SBR) and the influence of hydraulic retention time (HRT) on the SNAD process was investigated. Around 96% NH(4)(+)-N removal and 87% COD removal were observed at 9 d HRT. Marginal decreases in the removal efficiencies were observed when the HRT was reduced to 3d or the loading rate was increased by three times. On the other hand, a drastic decrease in NH(4)(+)-N and COD removals were observed when the DO, pH and temperature were dropped shockingly. The response of the SNAD system towards the shock in substrate loading and operating conditions was evaluated by sensitivity index. Finally, the extent of total nitrogen (TN) removal by partial nitrification with anammox and denitrification was modeled using stoichiometric relationship. Modeling results indicated a TN removal of 85-87% by anammox with partial nitrification and 7-9% by denitrification.
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http://dx.doi.org/10.1016/j.biortech.2010.11.024DOI Listing
May 2011

Simultaneous sulfate reduction and copper removal by a PVA-immobilized sulfate reducing bacterial culture.

Bioresour Technol 2010 Jun 12;101(12):4354-61. Epub 2010 Feb 12.

Institute of Environmental Engineering, National Chiao Tung University, 1001, University Road, Hsinchu, Taiwan.

The effect of a sulfate reducing bacteria immobilized in polyvinyl alcohol (PVA) on simultaneous sulfate reduction and copper removal was investigated. Batch experiments were designed using central composite design (CCD) with two parameters, i.e. the copper concentration (10-100mg/L), and the quantity of immobilized SRB in culture solution (19-235 mg of VSS/L). Response surface methodology (RSM) was used to model the experimental data, and to identify optimal conditions for the maximum sulfate reduction and copper removal. Under optimum condition, i.e. approximately 138.5mg VSS/L of sulfate reducing bacteria immobilized in PVA, and approximately 51.5mg/L of copper, the maximum sulfate reduction rate was 1.57 d(-1) as based on the first-order kinetic equation. The data demonstrate that immobilizing sulfate reducing bacteria in PVA can enhance copper removal and the resistance of the bacteria towards copper toxicity.
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http://dx.doi.org/10.1016/j.biortech.2010.01.094DOI Listing
June 2010

Degradation of carbofuran in aqueous solution by ultrasound and Fenton processes: Effect of system parameters and kinetic study.

J Hazard Mater 2010 Jun 1;178(1-3):320-5. Epub 2010 Feb 1.

Department of Environmental Engineering and Health, Yuanpei University, Hsinchu, Taiwan.

To establish an efficient oxidation process for carbofuran degradation, the effects of some operating parameters such as dosages of H(2)O(2), Fe(2+) and initial carbofuran concentrations were observed during carbofuran degradation by the ultrasound process, Fenton process and a combined ultrasound/Fenton process. The degradation kinetics of carbofuran was also examined based on the experimental data. The results show that more than 99% of the carbofuran was degraded by the ultrasound/Fenton process within short reaction time periods. Increased dosages of H(2)O(2) and Fe(2+) enhanced the degradation of carbofuran in the ultrasound and Fenton oxidation processes, but initial carbofuran concentrations decreased carbofuran degradation in both the Fenton and ultrasound/Fenton processes. The degradation kinetics of carbofuran by the three oxidation processes was found to be in accordance with first-order reaction kinetics. The results provide fundamental information about the treatment of carbofuran wastewater and/or other pesticides by the ultrasound/Fenton oxidation process.
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http://dx.doi.org/10.1016/j.jhazmat.2010.01.081DOI Listing
June 2010

Co-existence of anammox and denitrification for simultaneous nitrogen and carbon removal--Strategies and issues.

J Hazard Mater 2010 Jun 21;178(1-3):1-9. Epub 2010 Jan 21.

Institute of Environmental Engineering, National Chiao Tung University, 1001, University Road, Hsinchu City 30010, Taiwan.

The discovery of anaerobic ammonium oxidation (anammox) has greatly improved the understanding of the nitrogen cycle. Anammox provides great promise for the removal of nitrogen from wastewater, containing high concentration of ammonium. However, the presence of organic carbon is considered as unfavorable to this autotrophic process, i.e. anammox. Most of the real wastewaters contain both organic carbon and nitrogen. Under this circumstance, several processes have been established primarily for the complete removal of organic carbon. Subsequently, the wastewater containing no or low organic carbon and nitrogen is treated via a variety of nitrogen removal processes. The co-existence of anammox and denitrification could be useful for the simultaneous removal of nitrogen and organic carbon in a single system rather than a sequential chain of treatment. This review addresses the microbiology, strategies, consequences and the future research challenges in the co-existence of anammox and denitrification.
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http://dx.doi.org/10.1016/j.jhazmat.2010.01.077DOI Listing
June 2010

Co-composting of green waste and food waste at low C/N ratio.

Waste Manag 2010 Apr 19;30(4):602-9. Epub 2010 Jan 19.

Institute of Environmental Engineering, National Chiao Tung University, 1001, University Road, Hsinchu City 30010, Taiwan.

In this study, co-composting of food waste and green waste at low initial carbon to nitrogen (C/N) ratios was investigated using an in-vessel lab-scale composting reactor. The central composite design (CCD) and response surface method (RSM) were applied to obtain the optimal operating conditions over a range of preselected moisture contents (45-75%) and C/N ratios (13.9-19.6). The results indicate that the optimal moisture content for co-composting of food waste and green waste is 60%, and the substrate at a C/N ratio of 19.6 can be decomposed effectively to reduce 33% of total volatile solids (TVS) in 12days. The TVS reduction can be modeled by using a second-order equation with a good fit. In addition, the compost passes the standard germination index of white radish seed indicating that it can be used as soil amendment.
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http://dx.doi.org/10.1016/j.wasman.2009.11.023DOI Listing
April 2010

Simultaneous partial nitrification, anaerobic ammonium oxidation and denitrification (SNAD) in a full-scale landfill-leachate treatment plant.

J Hazard Mater 2010 Mar 22;175(1-3):622-8. Epub 2009 Oct 22.

Institute of Environmental Engineering, National Chiao Tung University, 1001 University Road, Hsinchu 30010, Taiwan, ROC.

The occurrence of simultaneous partial nitrification, anaerobic ammonium oxidation and denitrification (SNAD) observed in a single partially aerated full-scale bioreactor treating landfill-leachate is reported in this paper. At present, the full-scale bioreactor is treating an average leachate flow of 304 m(3)d(-1) with a sludge retention time between 12 and 18d. The average COD, NH(4)(+)-N and NO(3)(-)-N concentrations at the upstream end of the bioreactor, i.e., influent, are 554, 634 and 3 mg L(-1), respectively; whereas no NO(2)(-)-N is detected in the influent. The percentage removals of COD and NH(4)(+)-N in the bioreactor were 28% and 80%, respectively. A nitrogen mass balance approach was adopted to analyze the performance of SNAD in the full-scale bioreactor. The total nitrogen (TN) removal by combined partial nitrification and anaerobic ammonium oxidation is 68% and the heterotrophic denitrification contributes to 8% and 23% of TN and COD removals, respectively. The red granule in the bioreactor was analyzed by using fluorescence in situ hybridization and polymerase chain reaction. The results of both analytical methods confirm the presence of anaerobic ammonium oxidizing bacteria as the predominant species along with other Planctomycete-like bacteria. Overall, the SNAD process offers the simultaneous removals of nitrogen and COD in the wastewater.
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http://dx.doi.org/10.1016/j.jhazmat.2009.10.052DOI Listing
March 2010